For Research Use Only. Not for use in diagnostic procedures. Others KAPA HiFi HotStart Uracil plus ReadyMix Kit RUO KAPA HiFi HotStart Uracil+ ReadyMix Kit SEQ-KAPA-0020 07 959 079 001 7 959 079 001 07959079001 7959079001 07959079001 KAPA HiFi HotStart Uracil+ Kit (250rxn) KAPA HiFi HotStart Uracil+ ReadyMix Kit 07613336111697 Reagents, kits KK2802 250 reactions Not Available 07 959 052 001 7 959 052 001 07959052001 7959052001 07959052001 KAPA HiFi HotStart Uracil+ Kit (50rxn) KAPA HiFi HotStart Uracil+ ReadyMix Kit 07613336111680 Reagents, kits KK2801 50 reactions Not Available KAPA HiFi HotStart is a B-family DNA polymerase, engineered to have increased affinity for DNA, without the need for accessory proteins or DNA-binding domains. The intrinsic high processivity of the enzyme results in significant improvement in yield, speed and sensitivity when compared with wild-type B-family DNA polymerases and polymerase blends. When used for the amplification of next-generation sequencing (NGS) libraries, KAPA HiFi HotStart DNA Polymerase exhibits high yields with minimal amplification bias and provides extremely uniform sequence coverage.The read-ahead function of proofreading DNA polymerases detects pro-mutagenic uracil residues in the template strand and prevents further strand extension, thereby reducing or completely inhibiting PCR amplification. In KAPA HiFi HotStart Uracil+ DNA Polymerase, this uracilbinding pocket is inactivated to enable the amplification of uracil-containing DNA. The enzyme has the same error rate and shows the same high yield, low GC-bias and coverage uniformity as the unmodified KAPA HiFi HotStart DNA Polymerase, rendering it particularly advantageous for applications employing bisulfite DNA conversion, which typically produces low concentrations of AT-rich DNA.KAPA HiFi HotStart Uracil+ ReadyMix (2x) is a ready-touse cocktail containing all components required for PCR, except primers and template. The ReadyMix contains 0.3 mM of each dNTP at 1X (dATP, dCTP, dGTP, dTTP), and does not contain dUTP.KAPA HiFi HotStart Uracil+ DNA Polymerase has 5’→3’ polymerase and 3’-5’ exonuclease (proofreading) activity, but no 5’→3’ exonuclease activity. The strong 3’→5’ exonuclease activity results in extremely high accuracy during DNA amplification. A proprietary antibody inactivates the polymerase until the first cycle of denaturation, minimizing spurious amplification products that may result from nonspecific priming events during reaction setup and initiation, and increasing overall reaction efficiency. en Amplification of bisulfite-converted DNASodium bisulfite treatment converts unmethylated cytosines to uracil, while methylated cytosines remain unconverted. Uracil is replaced by thymine during subsequent PCR amplification, allowing base- pair resolution of DNA methylation via sequencing. KAPA HiFi HotStart Uracil+ ReadyMix provides high yields and robust amplification of bisulfite-converted DNA.The KAPA HiFi HotStart Uracil+ ReadyMix Kit is especially well-suited for NGS applications, providing reduced amplification bias and increased amplification efficiency of bisulfite-converted libraries. Library amplification with KAPA HiFi HotStart Uracil+ ReadyMix provides dramatic improvements in coverage depth uniformity, and more complete representation across reference sequences.Amplification of damaged DNA samplesCytosine deamination occurs spontaneously over long periods of time, and more rapidly at elevated temperatures, and results in the accumulation of uracil in DNA and among free nucleotides. KAPA HiFi HotStart Uracil+ DNA Polymerase offers improved success rates than other proofreading enzymes when attempting high- fidelity amplification of damaged, uracil-containing DNA templates.Prevention of amplicon contamination with UDGKAPA HiFi HotStart Uracil+ DNA Polymerase readily incorporates dUTP during amplification, and can therefore be used in conjunction with uracil-DNA-glycosylase (UDG) to prevent carryover contamination. dUTP is included in PCRs to allow for the digestion of amplicons that may contaminate subsequent reactions with UDG prior to amplification. en

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